The Role of NDUFA4L2 in Clear Cell Renal Cell Carcinoma

NDUFA4L2 在透明细胞肾细胞癌中的作用

• 批准号: 10218022
• 负责人: Jaclyn Kubala
• 金额: $ 0.98万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2020-10-25
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10218022
• 关键词: Affect; American Cancer Society; Cancer Etiology; Cancer Patient; Cell Culture Techniques; Cell Proliferation; Cell Survival; Cells; Cessation of life; Characteristics; Clear Cell; Clear cell renal cell carcinoma; Complex; Data; Development; Diagnostic Factor; Disease; Doctor of Philosophy; Drug Delivery Systems; Encapsulated; Fibroblasts; Genes; Glycolysis; Human; Hypoxia Inducible Factor; Kidney; Knockout Mice; Lead; Lipids; Malignant Epithelial Cell; Malignant Neoplasms; Mediating; Mentors; Metabolic; Methods; Mitochondria; Modeling; Mus; NADH dehydrogenase (ubiquinone); Nanotechnology; Organ; Oxidative Phosphorylation; Patients; Pharmaceutical Preparations; Pharmacology; Phenotype; Play; Process; Production; Proliferating; Renal Cell Carcinoma; Renal carcinoma; Role; Site; Small Interfering RNA; Testing; Therapeutic; Time; Transcriptional Activation; Transgenic Organisms; Tumor Suppressor Proteins; United States; VHL gene; Work; cancer pharmacology; carcinogenesis; curative treatments; doctoral student; effective therapy; experimental study; genome-wide; in vivo; innovation; insight; interest; knock-down; loss of function; mouse model; nanoparticle; novel; novel therapeutics; overexpression; prenatal; programs; protein function; targeted treatment; therapeutic target; tool; tumor

ABSTRACT Kidney cancer is one of the ten most common cancers in the US and accounts for over 100,000 deaths each year. Over 80% of kidney cancers are classified as clear cell renal cell carcinoma (ccRCC), and insight as to how this disease develops is scarce. One characteristic of ccRCC is the overexpression of NADH dehydrogenase (ubiquinone) 1 alpha subcomplex, 4-like 2 (NDUFA4L2), which is overexpressed in over 90% of ccRCC patients. Additionally, NDUFA4L2 is necessary for cell proliferation and survival, therefore indicating its importance in the development of ccRCC. However, the function of NDUFA4L2 has yet to be elucidated. Previous studies have shown that NDUFA4L2 plays a role in the switch from mitochondrial oxidative phosphorylation to glycolysis, a metabolic shift that is observed in ccRCC. Additional studies suggested that NDUFA4L2 may be implicated in the inhibition of mitochondrial complex I activity. However, this proposed function has yet to be investigated in ccRCC. Functional studies on NDUFA4L2 in vivo are lacking due to the prenatal lethality of NDUFA4L2 knockout mice. The Gudas lab has generated a TRAnsgenic Cancer of the Kidney (TRACK) murine model in which NDUFA4L2 is overexpressed. Additionally, this novel murine model recapitulates the human ccRCC disease, as demonstrated by genome- wide profiling of the TRACK kidneys. Previous collaborative work between the Gudas lab and the Heller lab showed that mesoscale nanoparticles (MNPs) localize in the mouse kidneys up to 25 times more than they do in other organs. Furthermore, these MNPs localized more in the “clear” cells, or lipid-filled cells, of the TRACK kidneys relative to normal proximal tubule cells. Consequently, these MNPs present a novel method of delivering drugs, therapeutics, or other investigative tools to the site of disease in the renal carcinogenesis murine model. We therefore aim to utilize this innovative approach to investigate the function of NDUFA4L2 in ccRCC. Through the utilization of the TRACK mouse model and the kidney-targeting MNPs, we will: 1) elucidate the role of NDUFA4L2 on mitochondrial function, and 2) determine the knockdown efficacy of siNDUFA4L2-encapsulating MNPs in TRACK mice. In order to investigate the mitochondrial function of NDUFA4L2, we will analyze mitochondrial function, ATP production, ROS levels, and complex I activity in cultured human ccRCC cells and TRACK mice. MNPs encapsulating siRNA specific for NDUFA4L2 will then be injected into TRACK mice to investigate whether NDUFA4L2 knockdown can be successfully achieved using this novel method. The proposed studies will delineate the functions of NDUFA4L2 in ccRCC and elucidate a possible delivery method of NDUFA4L2 knockdown specific to the kidneys. Findings from this proposal will provide critical insight into the underlying processes of ccRCC and present potential therapeutic strategies.

摘要 肾癌是美国最常见的十种癌症之一，每种癌症造成的死亡人数超过10万人 年。超过80%的肾癌被归类为肾透明细胞癌(CcRCC)， 这种疾病是如何发展的，目前还很少见。慢性肾细胞癌的一个特征是NADH的过度表达。 脱氢酶(泛醌)1α亚复合体，4-样2(NDUFA4L2)，90%以上过表达 在慢性肾细胞癌患者中。此外，NDUFA4L2是细胞增殖和存活所必需的，因此 表明其在中国农村合作与发展委员会发展中的重要性。然而，NDUFA4L2的功能还有待于 已澄清。先前的研究表明，NDUFA4L2在线粒体的转换中发挥了作用 氧化磷酸化到糖酵解，在ccRCC中观察到的一种代谢转变。其他研究 提示NDUFA4L2可能参与了线粒体复合体I活性的抑制。然而， 这一拟议的功能尚未在ccRCC中进行研究。NDUFA4L2的体内功能研究 缺乏是由于NDUFA4L2基因敲除小鼠的产前致死性。古达斯实验室已经产生了一个 转基因肾癌(TRACK)小鼠模型中NDUFA4L2过表达。 此外，这个新的小鼠模型概括了人类的ccRCC疾病，如基因组所证明的- 对赛道肾脏进行了广泛的分析。古达斯实验室和海勒实验室之前的合作工作 研究表明，中尺度纳米颗粒(MNPs)在小鼠肾脏中的分布是它们的25倍 在其他器官中。此外，这些MNPs更多地定位于脑组织的“透明”细胞或充满脂质的细胞。 追踪相对正常近端小管细胞的肾脏。因此，这些MNPs提供了一种新的方法 将药物、治疗药物或其他研究工具送到肾癌的发病部位 小鼠模型。因此，我们的目标是利用这种创新的方法来研究NDUFA4L2的功能 在ccrcc。通过利用跟踪小鼠模型和肾脏靶向MNPs，我们将：1) 阐明NDUFA4L2在线粒体功能中的作用，以及2)确定NDUFA4L2的敲除效率 SiNDUFA4L2-在跟踪小鼠体内包裹MNPs。为了研究线粒体的功能 NDUFA4L2，我们将分析线粒体功能、ATP产生、ROS水平和复合体I活性 培养人肾小管上皮细胞和Track小鼠。然后，封装NDUFA4L2特异性siRNA的mNPs将 注射入跟踪小鼠体内，研究能否成功实现NDUFA4L2基因敲除 使用这种新的方法。拟议的研究将描绘NDUFA4L2在ccRCC和ccRCC中的功能 阐明NDUFA4L2基因敲除的一种可能的肾脏特异性传递方法。由此得出的结论 该提案将提供对慢性肾细胞癌潜在过程的重要洞察，并提出潜在的治疗方法 战略。